Fine-tuning of a COI PCR-RFLP assay for fast genetic characterization of Spanish white-clawed crayfish

The white-clawed crayfish is endemic to western and southern Europe and its population has dramatically decreased in the last few decades. The latest reports on this species have shown that the genetic variability of Spanish populations displays a clear geographic pattern, with two main genetic groups across its range of distribution. To ensure the preservation of this endangered species, specimens for restocking purposes should be chosen, ideally, accordingly to their genetic characteristics. Here, we propose a PCR-RFLP assay developed for this purpose. Digestion of a fragment of the mitochondrial cytochrome oxidase subunit I gene – with two restriction enzymes, HpaI and CviAII - differentiates the most common haplotypes identified in Spain. This technique is a useful, low-cost and rapid method to facilitate the genetic characterization of Spanish white-clawed crayfish and, in doing so, to incorporate genetic information into conservation plans for this species.